Diagnosis of goose circovirus infection in Hungarian geese samples using polymerase chain reaction and dot blot hybridization tests

Abstract

A polymerase chain reaction (PCR) and dot blot hybridization (DBH) test have been developed for the diagnosis of infection by a novel circovirus of geese (GoCV). These tests were applied to samples of bursae of Fabricius from sick and dead birds from commercial goose farms in Hungary. In this second report of the occurrence of circovirus infection in diseased geese, 103 of 214 (48.1%) and 37 of 150 (24.6%) birds, and 49 of 76 (64.5%) and 18 of 76 (23.7%) flocks were positive by PCR and DBH respectively. The sensitivity of the PCR test was such that 0.10 fg of virus DNA was detectable. The DBH test was less sensitive, only detecting larger amounts (40 pg) of DNA, but was used as a semi-quantitative method for detecting the presence of virus. The incidence of infection was affected by factors such as the age of the birds and rearing methods.

Circovirus-infected geese studied byin situhybridization

“…These infections are now widely accepted as being highly significant, and therefore similar findings in geese raise the possibility that GoCV infection may well have clinicopathological significance. Since it has recently been shown that circovirus infection is widespread in farmed geese (Ball et al ., 2004), there is clearly a need for further work to accurately establish the significance of this infection. In conclusion, a sensitive and specific method of detecting goose circovirus DNA in formalin-fixed paraffin-embedded tissues has been developed.…”

“…In a recent study, Hungarian geese flocks diagnosed as infected by polymerase chain reaction (PCR) or dot blot hybridization were compared with non-infected flocks. With the exception of circovirus inclusions, which were found in the BF of birds in three infected flocks, no particular clinical syndrome or laboratory findings could be specifically associated with circovirus infection (Ball et al ., 2004). Thus, there is a need for further studies to establish the role, if any, of circoviruses in disease in geese.…”

Diagnosis of duck circovirus infections by conventional and real-time polymerase chain reaction tests

“…The PCR products were separated by electrophoresis on a 2% (w/v) of agarose gel and the DNA bands were stained with ethidium bromide (0.5 mg/ml) and visualized using ultraviolet light. The specificity of the test was evaluated by applying the PCR test to known GoCV-positive and PiCV-positive samples of BF (Todd et al ., 2002;Ball et al ., 2004), respectively. SYBR Green real-time PCR test.…”

“…This paper describes the first development of a PCR test for diagnosing DuCV infections and its use in assessing the prevalence of circovirus infections in dead and sick ducks. In the absence of serological tests, PCR has proved to be a specific and sensitive method for diagnosing avian circovirus infections (Ypelaar et al ., 1999;Todd et al ., 2002;Ball et al ., 2004). However, its ability to detect circovirus DNA in clinically normal birds, for example pigeons (Todd et al ., 2002), suggests that while it may be useful for diagnosing circovirus infections, PCR may be too sensitive to use as a diagnostic of disease.…”

“…However, its ability to detect circovirus DNA in clinically normal birds, for example pigeons (Todd et al ., 2002), suggests that while it may be useful for diagnosing circovirus infections, PCR may be too sensitive to use as a diagnostic of disease. This has prompted speculation that, in terms of clinical severity, the effects of circovirus infection may depend on virus load (Todd et al ., 2002;Ball et al ., 2004). With the aim of investigating the circovirus loads in infected ducks, this present paper also reports the development of a real-time PCR test, based on SYBR Green chemistry, and its application to selected BF samples.…”